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PART I CRYSTAL STRUCTURE OF A DIMERIZATION DOMAIN OF DROSOPHILA CAPRIN. PART II CHARACTERIZATION OF TWO CAS13B CRISPR-CAS SYSTEMS FROM PORPHYROMONAS GINGIVALISZhu, Jiang 01 May 2018 (has links) (PDF)
PART I: CRYSTAL STRUCTURE OF A DIMERIZATIO DOMAIN OF DROSOPHILA CAPRIN Drosophila Melanogaster Caprin (dCaprin) shares conversed HR1 domain with Caprin protein family members, which are RNA binding proteins that play critical roles in many important biological processes, such as synaptic plasticity, stress response, innate immune response and cellular proliferation. One of the Caprin protein family members, Caprin-1, is involved in the pathway of several human diseases, including breast cancer, neurodegenerative disorders, osteosarcoma, hearing loss, and viral infection. The functions of Caprin protein relies on their molecular interactions. Several direct interactions have been established between Caprin-1 and the Fragile X mental retardation protein (FMRP), Ras-GAP SH3 domain-binding protein 1 (G3BP1), and the Japanese encephalitis virus (JEV) core protein. We have determined the crystal structures of a fragment (residues 187-309) of Drosophila Melanogaster Caprin (dCaprin), which mediates homodimerization through a substantial interface created by a mainly alpha-helical fold. A larger hollow surface is created by homodimerization suggesting a protein binding groove. The FMRP binding should not affect dCaprin homodimerization for an integral alpha-helix in the dimeric dCaprein which formed by the FMRP interacting sequence motif. PART II: CHARACTERIZATION OF TWO TYPE VI-B CRESPR SYSTEMS: PGI5CAS1B AND PGI8CAS13B WHICH EFFECTOR PROTEINS ARE CAPABLE OF PROCESSING PRE-CRRNA INTO MATURE CRRNA CRISPR-Cas adaptive immune system protects microorganism from foreign nucleic acids invasion through endonucleases activity guided by RNA, which system has turned to a powerful genome editing tool applied to a multifold species, ranging from bacteria to human. Pgi5Cas13b and Pgi8Cas13b are identified by a computational sequence database mining approach, the CRISPR arrays lack of Cas1 and Cas2 encoding genes but contain a large candidate effector protein around 1,200 amino acids. They can be potentially classified as subtype VI-B CRISPR-Cas systems. We characterized the mature crRNA for Pgi5Cas13b and Pgi8Cas13b via Northern blot and small RNA sequencing. By EMSA (Electrophoretic mobility shift assay) experiments, we identified the binding constant between Pgi5Cas1b/Pgi8Cas13b and their corresponding crRNAs. The CRISPR loci of two of the Cas13b systems were cloned in pACYC vector and expressed in E. coli cells. Small RNAs were extracted and characterized by Northern Blotting and NGS (small RNA-seq) methods. The NGS results revealed the exact sequences of the crRNAs, which show a few new features not previously observed in other systems, including the longest spacer-derived sequences (32 and 31 nt), spacer-derived sequences flanking both ends of the full DR-derived sequence. The results also indicate different rules of pre-crRNA processing by the Pgi5Cas13b and Pgi8Cas13b systems. The characterization of these CRISPR systems extends the application of CRISPR based genome editing tools and promotes the development of single transcript manipulation tools.
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Characterization of Fluorescent Nanodiamonds containing Nitrogen-Vacancy and Silicon-Vacancy Color Centers as Produced by Pulsed Laser Ablation in Liquid ConfinementPiccoli, Alessandro 27 February 2024 (has links)
Nanodiamonds are a promising platform for quantum technologies due to the combinations of their inherent properties and the properties of the fluorescent color centers hosted in diamond. They can be employed as quantum sensing devices with spatial resolution in the range of the nanometer and capable of withstanding harsh conditions while also being biocompatible, allowing applications with sensitive biological systems; but they also find application in quantum computing and photonics fields. For all these applications the central features are the properties of the photoluminescent
color centers employed, the color centers on which this thesis is focused are the Nitrogen-Vacancy (NV) and Silicon-Vacancy (SiV) centers of diamond. Both centers are of high interest due to spin dependent properties of their
fluorescent emission which can be accessed at room temperature. The development of quantum technologies based on such fluorescent nanodiamonds is stifled by the the lack of production techniques that can be easily scaled to industrial levels. In fact most of the more prominent techniques found in literature exhibit drawbacks both in terms of control of particle properties and of scalability. This thesis focuses on the synthesis of nanodiamonds by Pulsed Laser Ablation in Liquid, with particular interest in the possibility of producing continuously nanodiamonds containing NV and SiV centers. For the NV center the technique of choice have been Pulsed Laser Ablation in liquid nitrogen focusing on the yield of the process as the technique has already been experimentally validated. For the SiV centers the ablation process was performed in water and the graphite precursor have been substituted for a composite graphite and silicon carbide precursor.
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Contextualisation & Capture de Gestuelles Utilisateur : Contributions à l'Adaptativité des Applications Interactives Scénarisées / Contextualization & User Body Language Capture : Contribution to the Adaptivity of Interactive ApplicationsPicard, François 11 July 2011 (has links)
Depuis 50 ans, une évolution permanente de l’interaction homme-machine a permis qu’aujourd’hui, nous tendions vers des systèmes temps réel proposant une interaction simple et intuitive à l’utilisateur et s’adaptant automatiquement à l’activité observée et interprétée. L’utilisateur peut dorénavant interagir avec un système informatique, volontairement ou de manière non consciente, par le biais de plusieurs modalités, comme il ferait dans la vie courante. Actuellement, les systèmes les plus développés sont ceux permettant à l’utilisateur d’interagir par le biais de gestuelles, qu’elles soient explicites ou implicites. Ces systèmes sont intégrés de plus en plus dans notre environnement, la plupart du temps invisibles pour nous, et s’adaptent en fonction d’un scénario qui définit les objectifs que nous devons atteindre.Plusieurs domaines d’application sont le cadre du développement de tels systèmes, comme celui du jeu vidéo ou encore de la surveillance vidéo. Nous proposons, dans ces travaux de thèse, l’architecture d’un système interactif, sur lequel s’exécute une application scénarisée de type jeu vidéo. L’interactivité est alimentée par les gestuelles du corps d’un unique utilisateur, en temps réel et de manière non invasive. Le système réagit également à divers événements prenant place au sein de la scène réelle, qui sont dus au dynamisme de cette dernière et dont l’utilisateur n’est pas directement responsable (changement d’éclairage, présence de spectateurs, etc.). L’utilisateur est immergé au sein d’un environnement virtuel qui traduit la réponse interactive du système. Ce dernier répond également, en temps réel, à l’activité observée et interprétée, de manière adaptative, adaptant à la fois sa réponse interactive et son fonctionnement global.Nos travaux se basent sur l’hypothèse que l’activité observée au sein de la scène est caractérisée par le contexte d’interaction qui l’englobe. Notre système reconnaît ainsi l’activité en modélisant le contexte d’interaction au sein duquel elle prend place. Notre contribution principale se traduit donc par l’introduction de la notion de contexte au sein des processus interactif et adaptatif. La modélisation du contexte d’interaction que nous proposons nous a servi de base pour celle du scénario d’une application. La gestion et l’analyse de ce contexte au cours de l’interactivité permettent au système d’interpréter l’activité observée et de paramétrer les différents mécanismes adaptatifs qui en découlent. L’activité est capturée et codifiée par un système étendu, adopté en accord avec le cadre industriel de cette thèse (convention CIFRE avec la société XD Productions). Initialement dédié à la capture, invasive et en milieu contrôlé, des mouvements de l’utilisateur, nos travaux ont permis l’augmentation du processus vers une capture plus générale de l’activité globale,de manière non invasive et en environnement dynamique. Enfin, nous avons développé une application immergeant l’utilisateur au sein d’une simulation virtuelle d’entrainement au tennis. Par le biais d’études de cas extraites de ce scénario, nous avons implémenté les processus interactif et adaptatif prenant place entre le système et l’utilisateur. L’adaptativité du système, supportée par le scénario de l’application, est concrétisée par la mise en place de mécanismes spécifiques à tous les niveaux de l’architecture, en fonction de l’activité observée et interprétée. Nous mettons en évidence un ensemble de boucles logicielles, appelées « boucles vertueuses », générées par l’accumulation des effets d’adaptation du système et améliorant en permanence l’interactivité. Les perspectives à ces travaux de thèse concernent la normalisation et la gestion haut niveau de notre modèle de contexte, la complexification de notre scénario et l’amélioration de notre système de capture dans le cadre de nouvelles applications scénarisées. / For 50 years, human-computer interaction has evolved permanently. Nowadays, real-time systems offer the user simple and intuitive interaction and adapt automatically to the observed and interpreted activity. From now on, the user can interact with computer systems, voluntarily or unconsciously, by several modalities, as he would do it in his everyday life.Currently, the most developed systems are the ones allowing the user to interact by his explicit or implicit body language. These systems are more and more integrated in our environment, are invisible, and adapt according to the scenario that defines the objectives the user has to achieve. Several application fields are the development frameworks of such systems, like video games or video surveillance. We define in this work the architecture of an interactive system, upon which runs a scripted application such as a video game. The interactivity is fed by an only user body language, in real-time and without markers. The system reacts as well to various events taking place within the real scene. These events aredue to the scene dynamism for which the user is not directly responsible (illumination changes, spectators, etc.).The user is immersed within a virtual environment that renders the system interactive response. The system responds adaptively as well, in real-time, to the observed and interpreted activity, adapting at the same time its interactive response and its global functioning. Our work is based on the following hypothesis: the activity, observed within the scene, is characterized by the interaction context that includes it. Thus, our system recognizes the activity by modelling the interaction context within which it takes place. Our main contribution is then the introduction of the notion of context within the interactive and adaptive processes. We use our interaction context modelling to model the application scenario. The context management and analysis during the interactivity allow the system to interpret the observed activity and to configure the different adaptive mechanisms which follow it. The activity is captured and codified by an extended system, chosen according to our work industrial framework (Industrial Convention of Formation by Research – CIFRE – with XDProductions). Initially dedicated to the invasive capture of the user movements in a controlled environment, our work enhances the process towards the general capture of the global activity, without markers and within an uncontrolled environment. Finally, we have developed an application immersing the user within a virtual simulation of tennis training. Thanks to different case studies extracted from this scenario, we have implemented the interactive and adaptive processes taking place between the system and the user. The system adaptivity, supported by the application scenario, is materialized by the implementation of specific mechanisms at every level of the architecture, according to the observed and interpreted activity. We have brought to light a set of software loops, called virtuous loops, generated by the accumulation of the system adaptation effects and improving the interactivity permanently. Our work perspectives concern the formalization and the high-level management of our context model, the complexity of our scenario and the improvement of our capture systemwithin the framework of new scripted applications.
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